SSR-labeled fingerprint of shiitake mushroom L9319 strain, and application thereof

A technology of shiitake mushrooms and strains, which is applied in the field of detection of shiitake mushroom strains, can solve the problems of affecting the enthusiasm of cultivation, affecting the rapid development of shiitake mushrooms, and the loss of mushroom farmers, and achieves good application prospects, short detection time and high accuracy.

Active Publication Date: 2014-11-12
SHANGHAI ACAD OF AGRI SCI
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Problems solved by technology

As far as the country is concerned, the phenomenon of "same species with different names and different species with the same name" in the cultivation of shiitake mushrooms has not only brought great losses to mushroom farmers, but also affected their enthusiasm for cultivation, and also greatly affected the rapid development of shiitake mushrooms in China. and, along with the appearance of large-scale industrialized cultivation methods, the requirements for the quality of the cultivated strains of shiitake mushrooms are getting higher and higher, and it is necessary to develop a more simple, fast and accurate strain identification technology to ensure that each batch of species is used. accurate

Method used

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  • SSR-labeled fingerprint of shiitake mushroom L9319 strain, and application thereof
  • SSR-labeled fingerprint of shiitake mushroom L9319 strain, and application thereof
  • SSR-labeled fingerprint of shiitake mushroom L9319 strain, and application thereof

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Experimental program
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Effect test

Embodiment 1

[0025] (1) Mycelium culture: transfer the mushroom strains to potato dextrose culture medium, culture in a shake flask at 23℃~25℃ in the dark, and collect the hyphae after 3d~4d;

[0026] (2) Genomic DNA extraction: use CTAB (hexadecyl trimethyl ammonium bromide) method to extract genomic DNA of the above-mentioned hyphae, detect the concentration and purity of total genomic DNA by ultraviolet spectrophotometry, and adjust the concentration of sample DNA to be consistent;

[0027] The genomic DNA extraction process of mycelium by CTAB method includes:

[0028] ① Grind the freeze-dried shiitake mushroom hyphae with liquid nitrogen into powder, add 2×CTAB extract preheated at 65°C, keep it at 65°C for 45-60 minutes, occasionally shake and mix;

[0029] ②Centrifuge at 12000rpm and 4℃ for 20min, and take the supernatant;

[0030] ③Add a mixture of phenol, chloroform and isoamyl alcohol at a volume ratio of 25:24:1 to the above supernatant, mix gently for 10 minutes, centrifuge at 12000 rpm,...

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Abstract

The invention relates to an SSR-labeled fingerprint of shiitake mushroom L9319 strain, and an application thereof. The fingerprint is composed of 7 pairs of SSR-labeled specific allelic fragments developed according to a shiitake mushroom genomic sequence. The application comprises the steps that: SSR-labeled amplification is carried out upon a shiitake mushroom strain; and an obtained band type is compared with the fingerprint. If the band type is consistent with the fingerprint, the strain is a shiitake mushroom L9319 strain. Compared with a conventional morphological detection method, an antagonistic test, and mushroom fruiting trials, the application provided by the invention has the advantages of short detection time, high accuracy, and good repeatability. In 25 collected nationally validated shiitake mushroom main cultivated strains of our nation, the fingerprint has shiitake mushroom L9319 strain specificity.

Description

Technical field [0001] The invention belongs to the field of detection of mushroom strains, and particularly relates to an SSR marker fingerprint of the mushroom L9319 strain and its application. Background technique [0002] The production of shiitake mushrooms in my country has grown rapidly from 19,500 tons in 1983 to 2.42 million tons in 2005, accounting for more than 70% of the total production of shiitake mushrooms in the world. It has rewritten the world’s edible fungi production rankings and has been shrinking with the top double Some experts predict that due to the rapid development of China's mushroom industry, it will become the world's highest production of edible mushrooms in the past 10 years. Chinese mushrooms have attracted the attention of the world mushroom industry with their amazing development speed, high-quality quality and low cost. Chinese mushrooms have become popular all over the world. [0003] The contribution rate of high-quality strains in the yield an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 章炉军谭琦宋春艳尚晓冬鲍大鹏杨慧张丹巫萍
Owner SHANGHAI ACAD OF AGRI SCI
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